scholarly journals Ingestion of Salmonella enterica Serotype Poona by a Free-Living Nematode, Caenorhabditis elegans, and Protection against Inactivation by Produce Sanitizers

2003 ◽  
Vol 69 (7) ◽  
pp. 4103-4110 ◽  
Author(s):  
Krishaun N. Caldwell ◽  
Barbara B. Adler ◽  
Gary L. Anderson ◽  
Phillip L. Williams ◽  
Larry R. Beuchat

ABSTRACT Free-living nematodes are known to ingest food-borne pathogens and may serve as vectors to contaminate preharvest fruits and vegetables. Caenorhabditis elegans was selected as a model to study the effectiveness of sanitizers in killing Salmonella enterica serotype Poona ingested by free-living nematodes. Aqueous suspensions of adult worms that had fed on S. enterica serotype Poona were treated with produce sanitizers. Treatment with 20 μg of free chlorine/ml significantly (α = 0.05) reduced the population of S. enterica serotype Poona compared to results for treating worms with water (control). However, there was no significant difference in the number of S. enterica serotype Poona cells surviving treatments with 20 to 500 μg of chlorine/ml, suggesting that reductions caused by treatment with 20 μg of chlorine/ml resulted from inactivation of S. enterica serotype Poona on the surface of C. elegans but not cells protected by the worm cuticle after ingestion. Treatment with Sanova (850 or 1,200 μg/ml), an acidified sodium chlorite sanitizer, caused reductions of 5.74 and 6.34 log10 CFU/worm, respectively, compared to reductions from treating worms with water. Treatment with 20 or 40 μg of Tsunami 200/ml, a peroxyacetic acid-based sanitizer, resulted in reductions of 4.83 and 5.34 log10 CFU/worm, respectively, compared to numbers detected on or in worms treated with water. Among the organic acids evaluated at a concentration of 2%, acetic acid was the least effective in killing S. enterica serotype Poona and lactic acid was the most effective. Treatment with up to 500 μg of chlorine/ml, 1% hydrogen peroxide, 2,550 μg of Sanova/ml, 40 μg of Tsunami 200/ml, or 2% acetic, citric, or lactic acid had no effect on the viability or reproductive behavior of C. elegans. Treatments were also applied to cantaloupe rind and lettuce inoculated with S. enterica serotype Poona or C. elegans that had ingested S. enterica serotype Poona. Protection of ingested S. enterica serotype Poona against sanitizers applied to cantaloupe was not evident; however, ingestion afforded protection of the pathogen on lettuce. These results indicate that S. enterica serotype Poona ingested by C. elegans may be protected against treatment with chlorine and other sanitizers, although the basis for this protection remains unclear.

Parasitology ◽  
2018 ◽  
Vol 146 (3) ◽  
pp. 314-320 ◽  
Author(s):  
Veeren M Chauhan ◽  
David I Pritchard

AbstractCaenorhabditis elegans is a free-living nematode that resides in soil and typically feeds on bacteria. We postulate that haematophagic C. elegans could provide a model to evaluate vaccine responses to intestinal proteins from hematophagous nematode parasites, such as Necator americanus. Human erythrocytes, fluorescently labelled with tetramethylrhodamine succinimidyl ester, demonstrated a stable bright emission and facilitated visualization of feeding events with fluorescent microscopy. C. elegans were observed feeding on erythrocytes and were shown to rupture red blood cells upon capture to release and ingest their contents. In addition, C. elegans survived equally on a diet of erythrocytes. There was no statistically significant difference in survival when compared with a diet of Escherichia coli OP50. The enzymes responsible for the digestion and detoxification of haem and haemoglobin, which are key components of the hookworm vaccine, were found in the C. elegans intestine. These findings support our postulate that free-living nematodes could provide a model for the assessment of neutralizing antibodies to current and future hematophagous parasite vaccine candidates.


2004 ◽  
Vol 67 (10) ◽  
pp. 2151-2157 ◽  
Author(s):  
STEPHEN J. KENNEY ◽  
GARY L. ANDERSON ◽  
PHILLIP L. WILLIAMS ◽  
PATRICIA D. MILLNER ◽  
LARRY R. BEUCHAT

Caenorhabditis elegans, a free-living nematode found in soil, has been shown to ingest human enteric pathogens, thereby potentially serving as a vector for preharvest contamination of fruits and vegetables. A study was undertaken to evaluate the efficacy of cleaners and sanitizers in killing Salmonella enterica serotype Newport in the gut of C. elegans. Adult worms were fed nalidixic acid–adapted cells of Escherichia coli OP50 (control) or Salmonella Newport for 24 h, washed, placed on paper discs, and incubated at temperatures of 4 or 20°C and relative humidities of 33 or 98% for 24 h. Two commercial cleaners (Enforce and K Foam Lo) and four sanitizers (2% acetic acid, 2% lactic acid, Sanova, and chlorine [50 and 200 μg/ml]) were applied to worms for 0, 2, or 10 min. Populations of E. coli and Salmonella Newport (CFU per worm) in untreated and treated worms were determined by sonicating worms in 0.1% peptone and surface plating suspensions of released cells on tryptic soy agar containing nalidixic acid. Populations of Salmonella Newport in worms exposed to 33 or 98% relative humidity at 4°C or 33% relative humidity at 20°C were significantly (P ≤ 0.05) lower than the number surviving exposure to 98% relative humidity at 20°C. In general, treatment of desiccated worms with cleaners and sanitizers was effective in significantly (P ≤ 0.05) reducing the number of ingested Salmonella Newport. Results indicate that temperature and relative humidity influence the survival of Salmonella Newport in the gut of C. elegans, and cleaners and sanitizers may not eliminate the pathogen.


2014 ◽  
Vol 60 (1) ◽  
Author(s):  
Romina E. D’Almeida ◽  
María R. Alberto ◽  
Phillip Morgan ◽  
Margaret Sedensky ◽  
María I. Isla

AbstractZuccagnia punctata Cav. (Fabaceae), commonly called jarilla macho or pus-pus, is being used in traditional medicine as an antiseptic, anti-inflammatory and to relieve muscle and bone pain. The aim of this work was to study the anthelmintic effects of three structurally related flavonoids present in aerial parts of Z. punctata Cav. The biological activity of the flavonoids 7-hydroxyflavanone (HF), 3,7-dihydroxyflavone (DHF) and 2´,4´-dihydroxychalcone (DHC) was examined in the free-living nematode Caenorhabditis elegans. Our results showed that among the assayed flavonoids, only DHC showed an anthelmintic effect and alteration of egg hatching and larval development processes in C. elegans. DHC was able to kill 50% of adult nematodes at a concentration of 17 μg/mL. The effect on larval development was observed after 48 h in the presence of 25 and 50 μg/mL DHC, where 33.4 and 73.4% of nematodes remained in the L3 stage or younger. New therapeutic drugs with good efficacy against drug-resistant nematodes are urgently needed. Therefore, DHC, a natural compound present in Z. punctata, is proposed as a potential anthelmintic drug.


Nematology ◽  
2005 ◽  
Vol 7 (5) ◽  
pp. 761-766 ◽  
Author(s):  
Nancy Lu ◽  
Rekha Balachandar

AbstractCaenorhabditis elegans is a free-living nematode cultured in an axenic medium, the Caenorhabditis elegans Maintenance Medium (CeMM), which contains B-vitamins, salts, amino acids, nucleic acid substituents, growth factors and glucose as an energy source. After initial experiments established that either pantothenate or pantethine would satisfy the vitamin B5 requirement in C. elegans, reproduction in the nematodes was measured in eight equimolar concentrations of calcium pantothenate, pantethine or coenzyme A. The optimal levels for pantothenate were found to be 7.5, 30 and 120 μg ml−1. The optimal levels for pantethine and coenzyme A were found to be 35 μg ml−1 and 100 μg ml−1, respectively. Among the three compounds, coenzyme A (at 100 μg/ml) supported a significantly greater population growth and, perhaps, is a more metabolically active form. Mild toxicity was demonstrated for pantothenate at 480μg ml−1, pantethine at 560 and 140 μg ml−1, and coenzyme A was found to exhibit toxicity at 410 and 1700 μg ml−1. Based on our results, we recommend that in the future the CeMM could be supplemented with pantothenate (7.5 μg ml−1) alone.


Parasitology ◽  
2004 ◽  
Vol 128 (S1) ◽  
pp. S49-S70 ◽  
Author(s):  
J. S. GILLEARD

There is increasing interest in the use of the free-living nematode Caenorhabditis elegans as a tool for parasitic nematode research and there are now a number of compelling examples of its successful application. C. elegans has the potential to become a standard tool for molecular helminthology researchers, just as yeast is routinely used by molecular biologists to study vertebrate biology. However, in order to exploit C. elegans in a meaningful manner, we need a detailed understanding of the extent to which different aspects of C. elegans biology have been conserved with particular groups of parasitic nematodes. This review first considers the current state of knowledge regarding the conservation of genome organisation across the nematode phylum and then discusses some recent evolutionary development studies in free-living nematodes. The aim is to provide some important concepts that are relevant to the extrapolation of information from C. elegans to parasitic nematodes and also to the interpretation of experiments that use C. elegans as a surrogate expression system. In general, examples have been specifically chosen because they highlight the importance of careful experimentation and interpretation of data. Consequently, the focus is on the differences that have been found between nematode species rather than the similarities. Finally, there is a detailed discussion of the current status of C. elegans as a heterologous expression system to study parasite gene function and regulation using successful examples from the literature.


2019 ◽  
Vol 6 (1) ◽  
pp. 29 ◽  
Author(s):  
Gastón Risi ◽  
Elena Aguilera ◽  
Enrique Ladós ◽  
Gonzalo Suárez ◽  
Inés Carrera ◽  
...  

Nematode parasites have a profound impact on humankind, infecting nearly one-quarter of the world’s population, as well as livestock. There is a pressing need for discovering nematicides due to the spread of resistance to currently used drugs. The free-living nematode Caenorhabditis elegans is a formidable experimentally tractable model organism that offers key advantages in accelerating nematicide discovery. We report the screening of drug-like libraries using an overnight high-throughput C. elegans assay, based on an automated infrared motility reader. As a proof of concept, we screened the “Pathogen Box” library, and identical results to a previous screen using Haemonchus contortus were obtained. We then screened an in-house library containing a diversity of compound families. Most active compounds had a conjugation of an unsaturation with an electronegative atom (N, O, or S) and an aromatic ring. Importantly, we identified symmetric arylidene ketones and aryl hydrazine derivatives as novel nematicides. Furthermore, one of these compounds, (1E,2E)-1,2-bis(thiophen-3-ylmethylene)hydrazine, was active as a nematicide at 25 µm, but innocuous to the vertebrate model zebrafish at 50 µm. Our results identified novel nematicidal scaffolds and illustrate the value of C. elegans in accelerating nematicide discovery using a nonlabor-intensive automated assay that provides a simple overnight readout.


Parasitology ◽  
1996 ◽  
Vol 113 (S1) ◽  
pp. S157-S173 ◽  
Author(s):  
R. E. Isaac ◽  
D. Macgregor ◽  
D. Coates

SUMMARYThe nematode nervous system employs many of the same neurotransmitters as are found in higher animals. The inactivation of neurotransmitters is absolutely essential for the correct functioning of the nervous system, In this article we discuss the various mechanisms used generally in animal nervous systems for synaptic inactivation of neurotransmitters and review the evidence for similar mechanisms operating in parasitic and free-living nematodes. The sequencing of the entireCaenorhabditis elegansgenome means that the sequence of nematode genes can be accessed from theC. elegansdatabase (ACeDB) and this wealth of information together with the increasing knowledge of the genetics of this free-living nematode will have great impact on all aspects of nematode neurobiology. The review will provide an insight into how this information may be exploited to identify and characterize target proteins for the development of novel anti-nematode drugs.


Parasitology ◽  
2017 ◽  
Vol 145 (8) ◽  
pp. 979-987 ◽  
Author(s):  
Gustavo Salinas ◽  
Gastón Risi

AbstractThe free-living nematode Caenorhabditis elegans is the simplest animal model organism to work with. Substantial knowledge and tools have accumulated over 50 years of C. elegans research. The use of C. elegans relating to parasitic nematodes from a basic biology standpoint or an applied perspective has increased in recent years. The wealth of information gained on the model organism, the use of the powerful approaches and technologies that have advanced C. elegans research to parasitic nematodes and the enormous success of the omics fields have contributed to bridge the divide between C. elegans and parasite nematode researchers. We review key fields, such as genomics, drug discovery and genetics, where C. elegans and nematode parasite research have convened. We advocate the use of C. elegans as a model to study helminth metabolism, a neglected area ready to advance. How emerging technologies being used in C. elegans can pave the way for parasitic nematode research is discussed.


2003 ◽  
Vol 66 (11) ◽  
pp. 1964-1971 ◽  
Author(s):  
KRISHAUN N. CALDWELL ◽  
GARY L. ANDERSON ◽  
PHILLIP L. WILLIAMS ◽  
LARRY R. BEUCHAT

Caenorhabditis elegans was studied to determine the potential role of free-living microbivorous nematodes as vectors for preharvest contamination of fruits and vegetables with foodborne pathogens. The propensity of C. elegans to be attracted to seven strains of Escherichia coli O157:H7, eight serotypes of Salmonella, six strains of Listeria monocytogenes, and cantaloupe juice was investigated. Twenty to 30 adult worms were placed on the surface of K agar midway between a 24-h bacterial colony and 10 μl of uninoculated tryptic soy broth (TSB) or cantaloupe juice positioned 1.5 cm apart. The numbers of nematodes that migrated to the colony, to the TSB, and to the cantaloupe juice within 5, 10, 15, and 20 min at 21°C were determined, and then the plates were incubated at 37°C for up to 7 days to determine the ability of C. elegans to survive and reproduce in bacterial colonies. The nematode was attracted to colonies of all test pathogens and survived and reproduced within colonies for up to 7 days. C. elegans was not attracted to cantaloupe juice. The potential of C. elegans to serve as a vector for the transport of Salmonella Poona to cantaloupe rinds was investigated. Adult worms that had been immersed in a suspension of Salmonella Poona were deposited 1 or 3 cm below the surface of soil on which a piece of cantaloupe rind was placed. The rind was analyzed for the presence of Salmonella Poona after 1, 3, 7, and 10 days at 21°C. The presence of Salmonella Poona was evident more quickly on rinds positioned on soil beneath which C. elegans inoculated with Salmonella Poona was initially deposited than on rinds positioned on soil beneath which Salmonella Poona alone was deposited. The time required to detect Salmonella Poona on rinds was longer when the rind was placed 3 cm above the inoculum than when the rind was placed 1 cm above the inoculum. Free-living nematodes may play a role in the preharvest dispersal of incidental human pathogens in soil to the surfaces of raw fruits and vegetables in contact with soil during development and maturation, as evidenced by the behavior of C. elegans as a test model.


1998 ◽  
Vol 336 (3) ◽  
pp. 545-550 ◽  
Author(s):  
Akram A. DA'DARA ◽  
Rolf D. WALTER

S-Adenosylmethionine decarboxylase (SAMDC) is a major regulatory enzyme in the polyamine biosynthesis and is considered a potentially important drug target for the chemotherapy of proliferative and parasitic diseases. To study regulatory mechanisms which are involved in the expression of SAMDC of the free-living nematode Caenorhabditis elegans, we have isolated the SAMDC gene and cDNA. Genomic Southern-blot analysis suggests that the C. elegans SAMDC is encoded by a single-copy gene which spans 3.9 kb and consists of six exons and five introns. The first two introns are located in the 5´-untranslated region (UTR). Analyses of the 5´-flanking region of the gene revealed several consensus sequences for the binding of different transcription factors such as CBP, AP2, cMyb, VPE2 and others. The C. elegans SAMDC mRNA possesses an open reading frame (ORF) which encodes a polypeptide of 368 amino acids, corresponding to a SAMDC proenzyme with a calculated molecular mass of 42141 Da. The active form of the C. elegans SAMDC is a heterotetramer, consisting of two subunits of 32 and 10 kDa derived from cleavage of the pro-enzyme. The SAMDC mRNA has an unusually long 5´-UTR of 477 nucleotides. This region has a small ORF which could encode a putative peptide of 17 residues. Moreover, the C. elegans SAMDC mRNA is trans-spliced with the 22 nucleotides spliced leader sequence at the 5´-end.


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